Figure 4.
ERp29 overexpression induces TNTs in an MSec-dependent manner. A, U2OS cells transiently expressing either EGFP-ERp29 or luminal EGFP (control vector) were treated with siRNAs against MSec or luciferase (control) and co-stained with anti-GFP antibody (green) and phalloidin conjugated with Alexa FluorTM 594 (red). Cells were imaged using confocal microscopy. Arrows indicate TNTs connecting neighboring cells. Scale bar, 20 μm. B, quantification of the number of TNTs per 100 cells from the confocal images. Data represent mean ± S.D. (error bars) based on three independent experiments, 100 cells counted per experiment (two-way ANOVA, Sidak's multiple-comparison test; *, p < 0.05; ****, p < 0.0001). C, efficiency of overexpression of EGFP-ERp29 and depletion of MSec analyzed by immunoblotting with their corresponding antibodies.